2005, 32 (3) : Acta Horticulturae Sinica 449453 AFL P (, 100081) : AFLP, 3 + 2, 48, 3 184DNA, 5613, 9118%, DNA :, AFLP, : ; ; ; AFLP; : S 63213: A : 05132353X (2005) 0320449205 AFL P Ana lysis of Genetic D iversity of Taro Germ pla sm s in Y unnan Prov ince Shen D i, Zhu Dewei, L i Xixiang, and Song J iangp ing ( Institute of V egetables and Flow ers, Chinese A cadem y of A gricultural Sciences, B eijing 100081, China) Abstract: The genetic diversity of 48 taro Colocasia escu lenta (L. ) Schottgermp lasm resources collected from Yunnan p rovince were analyzed by AFLP technique and 3 + 2p rimers labeled with fluores2 cence. 184 DNA bands were amp lified by three pairs of AFLP p rimers and 5613 polymorphic bands per pair of p rimer were p roduced on average. The percentage of polymorphic bands was 9118%. R ich genetic diversity of taro germp lasm was disp layed at DNA level in Yunnan p rovince. C luster analysis showed that the relationship between w ild germp lasm s and cultivars was far, and the classification result based on AFLP markers of culti2 vars was basically consistent w ith those on their morphological characters. Key words: Taro; Germp lasm resource; Genetic diversity; AFLP; C luster analysis Colocasia esculenta (L. ) Schott(A raceae),,, 53 1,,,,,, RAPD 24 AFLP, 48,, 1 111 481, 18, 8 (3671322252642), : 2004-09 - 08; : 2005-02 - 06 : 5(1516171837) 35 :
450 32 No. Local name 148 3 Table 1The or ig ina l informa tion of taro germ pla sm s Source Stolon Petiole color Color of leaf center 1 Baoshan Da Yutou Cultivar Absent Greenish white Purp le red 2 Yimen Q ingyu Cultivar Absent Green Greenish white 3 Maguan Yeyu W ild Present L ight purp le Purp le red 4 Laiyuhe Cultivar Absent Greyish white Greyish W hite 5 Hong Yu Cultivar Absent L ight purp le Purp le red 6 Ye Yu W ild Present L ight purp le Purp le red 7 Chengga Yeyu W ild Present L ight red Purp le 8 Heigeng Yu Cultivar Absent Dark purp le Purp le 9 Bai Yu Cultivar Absent Greenish white Greenish white 10 Kua mo mei( Bai Yu) Cultivar Absent Green Greenish white 11 Baigeng Ziyu Cultivar Present Purp le Purp le 12 Honggeng Ziyu Cultivar Absent L ight purp le Greenish white 13 L geng Yeyu W ild Present L ight purp le Purp le 14 Xiao L yu Cultivar Absent Greenish white Greenish white 15 Ye Yu Sem i2wild Present 16 Duoya L geng Yeyu Sem i2wild Present L ight purp le Purp le 17 Jiucheng W angenyu Sem i2wild Present Green Greenish white 18 Tuan Yu Sem i2wild Absent Greenish white Red spot 19 Tongbiguan W angenyu Cultivar Present Purp le spot Greenish white 20 Chushui Yu Cultivar Absent Red spot Purp le red 21 Hongya Yu Cultivar Absent L ight purp le Red spot 22 Husa Yeyu W ild Present Green Purp le 23 Hong Yu Cultivar Absent L ight red Purp le red 24 Rentou Yu Cultivar Absent L ight purp le Greenish white 25 Caiwen Yeyu W ild Present Green Purp le red 26 Shidian W angenyu W ild Present Green Purp le 27 Bai Yu Cultivar Absent Green Greenish white 28 Bai Yu Cultivar Absent Green Greenish white 29 LYu Cultivar Absent Green Red spot 30 Shui Yu Cultivar Absent L ight purp le Purp le red 31 D ishuiyu Cultivar Absent Greyish white Greyish white 32 Zi Yu Cultivar Absent Dark purp le Purp le red 33 Hongya Yu Cultivar Absent Greenish white Purp le red 34 Hong Yu Cultivar Absent Purp le red Purp le red 35 Hongxiang Yu Cultivar Absent Green Red Spot 36 Xanthosoma ( Zi Yu) Cultivar Absent Dark purp le Dark purp le 37 Xiao Bai Yu Sem i2wild Absent Greenish white Greenish white 38 Byong ma ne ( Hong Yu) Cultivar Absent Dark purp le Purp le 39 Byong ma a yo p iu ( Hong Yu) Cultivar Absent Purp le red Purp le red 40 Byong ma byong na ( Zi Yu) Cultivar Absent Dark purp le Purp le red 41 Huo pe ( Zi Yu) Cultivar Absent L ight purp le Purp le 42 Ye Yu W ild Present Green Purp le 43 Bu le ce ( Lu Yu) Cultivar Absent Greenish white Purp le 44 Bu le na ( Hei Yu) Cultivar Absent Dark purp le Purp le 45 B i ge a na ( Zi Yu) Cultivar Absent Purp le spot Purp le 46 Ku biu ( Hong Yu) Cultivar Absent L ight purp le Purp le red 47 Sangzangyu Cultivar Absent Green Purp le red 48 Hong Yu Cultivar Absent 3 3 The background information above was p rovided by the Kunm ing Institute of Botany, Chinese Academy of Sciences. 112D NA - DNA 5
3 : AFLP 451 113AFL P PCR DNA 5 114AFL P L i2cordna 4200 L, 6%, 012 mm, 1 TBE, 1L, MarkerL i2cor, () 50100145200204255300350364400460495500530565600 650 bp 115 1, 1, 0 AFLP 50350 bp, 350 bp50 bp, (p) : p = ( k /n) 100%, k ; n Statistica, Unweighted Pair2Group Average 2 211AFL P 469, 3 + 2, E2 AAE2ACE2TTE2TG ( 4 ) M47M48 M49M50M59M60M61M62 ( 8 ) 32,, M472E2AAM492E2AAM602E2TGM592E2TT M592E2TG 5 (2), 212AFL P 5 48 AFLP, DNA 100350 bp, M472E2AA (1) M492E2AAM602E2TG 3 DNA 3 184, 15 48 5613, 9118% (3), AFLP, 21348 2AFL P5 Table 2The ba se sequence of f ive prim er com b ina tion s Primer Combination M472E2AA M492E2AA M592E2TT M602E2TG M592E2TG for AFL P ana lysis Base sequence 5 2 GATGAGTCCTGAGTAACAA23, 5 2 GACTGCGTACCAATTCAA23 5 2 GATGAGTCCTGAGTAACAG23, 5 2 GACTGCGTACCAATTCAA23 5 2 GATGAGTCCTGAGTAACTA23, 5 2 GACTGCGTACCAATTCTT23 5 2 GATGAGTCCTGAGTAACTC23, 5 2 GACTGCGTACCAATTCTG23 5 2 GATGAGTCCTGAGTAACTA23, 5 2 GACTGCGTACCAATTCTG23 3348 Table 3The result am plified by 3 pr im er com b ina tion Primer combination in 48 taro accessions Total band number Polymorphic band number M472E2AA 80 73 91 M492E2AA 72 65 90 M602E2TG 32 31 97 / Total/Average 184 /6113 169 /5613 9118 Polymorphic percentage ( % ) 48AFLP 3184DNA,, 1013, 318 (2), 810, 48, 13, 431, 5, 483
452 32 F ig. 148 AFL P(M 472E2AA) 1Band ing pa ttern s of 48 taro ma teria ls am plif ied by the prim er com b ina tion M 472E2AA 2 2 355 14 31, 610, 140314: 1 112 10,,,, 8, 212, ( 1 ) (18) (20) (24) ; 2 1922 8, ; 3 944 6, 9 10, 5 4(16304144), ; 4 740 7,, 37 () 3 AFLP AFLP48,,, F ig. 248 AFL P 2D endrogram of taro germ pla sm ba sed on AFL P da ta unweighted pa ir2group average, euclidean distances
3 : AFLP 453 9118%, (Colocasia) 13 6, 8, 4 4,, AFLP,,,, AFLP,,, 4 10RAPD,, 7, AFLP 8 AFLP 396, DNA, DNA, 9 V iet 10 48,, AFLP, : 1.., 2001, ( ) : 412 Ye J Y. ( in Chinese) The cultivation, origination and distribution of hydrophily vegetables in China. Changjiang Vegetable, 2001, ( Supp lement) : 412 2,.., 1997, 26 (4) : Chen W B, Zhang GM. 421426 The types of esterase isozymes of taro in China and the classification of cultivar groups. Journal of Fujian Agricultural University, 1997, 26 (4) : 421426 ( in Chinese) 3Lebot V, A rakhya K M. Isozyme variation in taro (Colocasia esculenta) from A sia and Ocaeania. Euphytica, 1991, 56: 5566 4,,. RAPD., 2001, 16 (4) : Long W H, XuM H, Zhang Y H. Prelim inary studies on taro (Colocasia Schott) cultural University, 2001, 16 (4) : 274279 ( in Chinese) 274279 in Yunnan with RAPD technique. Journal of Yunnan Agri2 5. :. :, 2000. 1923, 39 Shen D. Analysis of genetic diversity in taro Colocasia esculenta (L. ) Schottgermp lasm in Yunnan p rovince: Master Degree D isserta2 tion. Beijing: Chinese Academy of Agricultural Science, 2000. 1923, 39 ( in Chinese) 6,.. :, 1979. 6879 W u Z Y, L i H. The p lant index of China. Beijing: Scientific Press, 1979. 6879 ( in Chinese) 7,,,,,. AFLP., 2004, 31 (3) : 309314 L i X X, Zhu D W, Du Y C, Shen D, Kong Q S, Song J P. Studies on genetic diversity and phylogenetic relationship of cucumber (Cucum is sativus L. ) germp lasm by AFLP technique. Acta Horticulturae Sinica, 2004, 31 (3) : 309314 ( in Chinese) 8,,,,,. AFLP., 2003, 30 (4) : Yi G J, Huo H Q, Chen D C, Huang Z R, Cai C H, Q iu Y P. Horticulturae Sinica, 2003, 30 (4) : 399403 ( in Chinese) 9,,.., 1999, 1: 399403 Studies on genetic relationship among L itchi varieties by using AFLP. Acta Pu Y D, Yang Y P, Xu J C. Germp lasm resources and utilization of taro in Yunnan. Crop Germplasm Resources, 1999, 1: 14 ( in Chinese) 10V iet Xuan Nguyen, H irom ichi Yoshino et al. Phylogenetic analyses of taro Colocasia esculenta (L. ) Schottand related species based on esterase isozyme. Scientific Reports of the Faculty of Agriculture Okayama University, 1998, 87: 133139 14